SNX3-retromer requires an evolutionary conserved MON2:DOPEY2:ATP9A complex to mediate Wntless sorting and Wnt secretion

Nat Commun. 2018 Sep 13;9(1):3737. doi: 10.1038/s41467-018-06114-3.

Abstract

Wntless transports Wnt morphogens to the cell surface and is required for Wnt secretion and morphogenic gradients formation. Recycling of endocytosed Wntless requires the sorting nexin-3 (SNX3)-retromer-dependent endosome-to-Golgi transport pathway. Here we demonstrate the essential role of SNX3-retromer assembly for Wntless transport and report that SNX3 associates with an evolutionary conserved endosome-associated membrane re-modelling complex composed of MON2, DOPEY2 and the putative aminophospholipid translocase, ATP9A. In vivo suppression of Ce-mon-2, Ce-pad-1 or Ce-tat-5 (respective MON2, DOPEY2 and ATP9A orthologues) phenocopy a loss of SNX3-retromer function, leading to enhanced lysosomal degradation of Wntless and a Wnt phenotype. Perturbed Wnt signalling is also observed upon overexpression of an ATPase-inhibited TAT-5(E246Q) mutant, suggesting a role for phospholipid flippase activity during SNX3-retromer-mediated Wntless sorting. Together, these findings provide in vitro and in vivo mechanistic details to describe SNX3-retromer-mediated transport during Wnt secretion and the formation of Wnt-morphogenic gradients.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism*
  • Animals
  • Biological Transport
  • Caenorhabditis elegans
  • Endosomes / metabolism*
  • Golgi Apparatus / metabolism*
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Membrane Transport Proteins / metabolism*
  • Mutation
  • Phenotype
  • Phospholipid Transfer Proteins / metabolism*
  • Protein Binding
  • Protein Domains
  • Proteomics
  • Proton-Translocating ATPases / metabolism*
  • RNA Interference
  • Sorting Nexins / metabolism*
  • Transgenes
  • Vesicular Transport Proteins / metabolism*
  • Wnt Proteins / metabolism*

Substances

  • DOP1B protein, human
  • MON2 protein, human
  • Membrane Transport Proteins
  • Phospholipid Transfer Proteins
  • SNX3 protein, human
  • Sorting Nexins
  • Vesicular Transport Proteins
  • Wnt Proteins
  • Green Fluorescent Proteins
  • Adenosine Triphosphatases
  • Proton-Translocating ATPases
  • ATP9A protein, human